Self-referenced axial chromatic dispersion measurement in multiphoton microscopy through 2-color third-harmonic generation imaging
Self-referenced axial chromatic dispersion measurement in multiphoton microscopy through 2-color third-harmonic generation imaging
复制标题
通过 2 色三次谐波生成成像在多光子显微镜中进行自参考轴向色散测量
DOI：
10.1002/jbio.201800071
复制
发表时间：
2018
影响因子：
2.8
通讯作者：
Wang Ke
中科院分区：
文献类型：
--
作者：
Du Yu;Zhuang Ziwei;He Jiexing;Liu Hongji;Qiu Ping;Wang Ke
多光子显微镜利用可调激发光，广泛用于亚细胞分辨率的生物结构成像。轴向色散，存在于几乎每一个透射光学系统，包括多光子显微镜，导致焦点（和所得图像）平面分离。在这里，我们实验证明了一种技术来测量轴向色散的多光子显微镜，使用同时2色三次谐波产生成像激发的2色孤子源与可调波长分离。我们的技术是自参考的，消除了使用单色可调激发光时的潜在测量误差，因为单色可调激发光需要机械平移台往复运动。使用这种技术，我们证明了测量轴向色散与2个不同的物镜在多光子显微镜。在生物样品中的进一步测量还表明，这种轴向色散与2色成像相结合，可以为2个不同轴向平面的同时成像提供机会。
With tunable excitation light, multiphoton microscopy is widely used for imaging biological structures at subcellular resolution. Axial chromatic dispersion, present in virtually every transmissive optical system including the multiphoton microscope, leads to focal (and the resultant image) plane separation. Here, we experimentally demonstrate a technique to measure the axial chromatic dispersion in a multiphoton microscope, using simultaneous 2‐color third‐harmonic generation imaging excited by a 2‐color soliton source with tunable wavelength separation. Our technique is self‐referenced, eliminating potential measurement error when 1‐color tunable excitation light is used which necessitates reciprocating motion of the mechanical translation stage. Using this technique, we demonstrate measured axial chromatic dispersion with 2 different objective lenses in a multiphoton microscope. Further measurement in a biological sample also indicates that this axial chromatic dispersion, in combination with 2‐color imaging, may open up opportunity for simultaneous imaging of 2 different axial planes.